Details zur Publikation

Referenztyp Zeitschriften
DOI / URL Link
Titel (primär) Standardized peptidome profiling of human cerebrospinal fluid by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
Autor Bruegel, M.; Planert, M.; Baumann, S.; Focke, A.; Bergh, F.T.; Leichtle, A.; Ceglarek, U.; Thiery, J.; Fiedler, G.M.;
Journal / Serie Journal of Proteomics
Erscheinungsjahr 2009
Department METABOX; PROTEOM;
Band/Volume 72
Heft 4
Sprache englisch;
Keywords Cerebrospinal fluid; MALDI-TOF; Mass spectrometry; Peptidome; Proteome; Standardization
Abstract Peptidome profiling of human cerebrospinal fluid (CSF) is a promising tool to identify novel disease-associated biomarkers. Our aim was to develop a standardized protocol for reproducible peptidome profiling of CSF using magnetic bead (MB) separation followed by MALDI-TOF MS. Peptidome fractionation and profiling of CSF were performed using MBs with different surface functionalities. We investigated exogenous variables (storage conditions, freeze-thaw-cycles) and endogenous interferences (albumin, immunoglobulin, blood, leukocytes) in pooled CSF samples. We detected approximately 500 signals with an S/N ratio > 10 and an overlap frequency of about 40% in non-pathological CSF. Within- and between-day imprecisions in relative signal intensities ranged from 3 to 28% and 7 to 47%, respectively. CSF storage at room temperature for up to 6 h and at 4 °C for up to 3 days did not significantly influence the mass spectra. Consecutive freeze-thaw-cycles significantly affected the mass spectra. High albumin and immunoglobulin content altered the CSF preparation using MB-HIC C8 beads. Blood contamination showed no effect on mass spectra up to a hemoglobin concentration of 0.075 µmol/L. The presence of leukocytes up to a cell number of 30 Mpt/L did not affect mass spectra. Our reliable pretreatment protocol allows standardization of preanalytical modalities and thereby enables reproducible peptidome profiling of human CSF using MB separation followed by MALDI-TOF MS.
ID 9838
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=9838
Bruegel, M., Planert, M., Baumann, S., Focke, A., Bergh, F.T., Leichtle, A., Ceglarek, U., Thiery, J., Fiedler, G.M. (2009):
Standardized peptidome profiling of human cerebrospinal fluid by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
J. Proteomics 72 (4), 608 - 615